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Tiêu đề New insight into silica deposition in horsetail (Equisetum arvense)
Tác giả Chinnoi Law, Christopher Exley
Người hướng dẫn Christopher Exley
Trường học Keele University
Chuyên ngành Plant Biology
Thể loại Research article
Năm xuất bản 2011
Thành phố Staffordshire
Định dạng
Số trang 9
Dung lượng 6,69 MB

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All of the major sites of silica deposition in horsetail mimicked sites and structures where the hemicellulose, callose is known to be found and these serendipitous observations of the c

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R E S E A R C H A R T I C L E Open Access

New insight into silica deposition in horsetail

(Equisetum arvense)

Chinnoi Law and Christopher Exley*

Abstract

Background: The horsetails (Equisetum sp) are known biosilicifiers though the mechanism underlying silica

deposition in these plants remains largely unknown Tissue extracts from horsetails grown hydroponically and also collected from the wild were acid-digested in a microwave oven and their silica‘skeletons’ visualised using the fluor, PDMPO, and fluorescence microscopy

Results: Silica deposits were observed in all plant regions from the rhizome through to the stem, leaf and spores Numerous structures were silicified including cell walls, cell plates, plasmodesmata, and guard cells and stomata at varying stages of differentiation All of the major sites of silica deposition in horsetail mimicked sites and structures where the hemicellulose, callose is known to be found and these serendipitous observations of the coincidence of silica and callose raised the possibility that callose might be templating silica deposition in horsetail Hydroponic culture of horsetail in the absence of silicic acid resulted in normal healthy plants which, following acid digestion, showed no deposition of silica anywhere in their tissues To test the hypothesis that callose might be templating silica deposition in horsetail commercially available callose was mixed with undersaturated and saturated solutions

of silicic acid and the formation of silica was demonstrated by fluorimetry and fluorescence microscopy

Conclusions: The initiation of silica formation by callose is the first example whereby any biomolecule has been shown to induce, as compared to catalyse, the formation of silica in an undersaturated solution of silicic acid This novel discovery allowed us to speculate that callose and its associated biochemical machinery could be a missing link in our understanding of biosilicification

Keywords: Biosilicification, biogenic silica, silicic acid, horsetails, callose, PDMPO, fluorescence, acid digestion

Background

Silicon is the second most abundant element of the

Earth’s crust after oxygen and, perhaps surprisingly, its

essentiality in biota remains equivocal [1] The difficulty

in ascribing true biochemical essentiality to silicon

prob-ably emanates from a lack of demonstration of any

sili-con-requiring biochemistry and specifically Si-C,

Si-O-C, Si-N, et c bonds in any form of extant life [2]

How-ever, in spite of such limitations the essentiality of

sili-con in plants remains the subject of rigorous debate

[3,4] as do elaborations of the underlying mechanisms

Biosilicification was recently defined as’the movement of

silicic acid from environments in which its concentration

does not exceed its solubility (< 2 mM) to intracellular

or systemic compartments in which it is accumulated for subsequent deposition as amorphous hydrated silica’ [5] and a number of plants are known biosilicifiers [4] One

of the best known of these are the horsetails, Equisetum sp., and silica deposition in the tissues of these plants has been studied extensively [6-12], perhaps the seminal work in the field being carried out by Perry and Fraser [13] In this work scanning and transmission electron microscopy was used to illuminate the elaborate and detailed micromorphology and ultrastructure of silicas extracted from different regions of the horsetail, Equise-tum arvense The images of silicified stomata and other silica sculptures are truly breathtaking and the level of organisation of silica in the tissues prompted the authors to speculate that’the silica acts as an in vivo stain, faithfully reproducing the organic matrix skeleton

at the microscopic and macroscopic levels without stain-ing’ Perry and Lu (1992) suggested that the organic

* Correspondence: c.exley@chem.keele.ac.uk

The Birchall Centre, Lennard-Jones Laboratories, Keele University,

Staffordshire, ST5 5BG, UK

© 2011 Law and Exley; licensee BioMed Central Ltd This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and

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matrix in question might be made from polymers of

car-bohydrates, for example, cellulose [14], and this

sugges-tion was reinforced recently by Fry and colleagues who

speculated that the hemicellulose, callose, in horsetail

cell walls might be a potential site of silica deposition

[15] Many different biomolecules, often having

origin-ally been extracted from biogenic silica, have been

shown to accelerate or catalyse silica deposition in

satu-rated solutions of silicic acid [16] However, biosilicifiers,

such as horsetails, harvest silicic acid from solutions

which are far from saturation and deposit it as

amor-phous hydrated silica and it is the elucidation of this

mechanism which remains the ‘Holy Grail’ of biological

silicification research [5]

Herein we have taken inspiration from the work of

Perry and Fraser [13] on horsetail and we have used

fluorescence microscopy to investigate biosilicification in

horsetail and to identify the organic matrix involved in

templating silica deposition in this plant

Results

PDMPO as a fluorescent marker of biosilicification

Microwave-assisted acid digestion of horsetail, either

grown hydroponically in the presence of silicic acid or

in plants collected from the wild, resulted in silica

deposits and‘skeletons’ which were successfully labelled

with the fluor PDMPO Silica was identified in acid

digests of all areas of the plant from the rhizome

through to spores in the cone There were no

structu-rally-distinct silica skeletons in the root, only what

appeared as diffuse deposits of siliceous materials

(Fig-ure 1a) Silica skeletons of basal stem showed

epider-mal-like cells, 30-40 μm wide and 100-300 μm long,

with heavily silicified cell walls and approximately

equi-distant punctate deposits of silica within the walls which

were suggestive of the expected locations of

plasmodes-mata Each‘silica cell’ included an amorphous, spherical

silica deposit between 10 and 20μm in diameter which

had the appearance of a nucleus or vesicle There were

also occasional heavily silicified (as indicated by an

enhanced fluorescence) skeletons of stomata,

approxi-mately 40μm wide and 70 μm long, which appeared to

be at various stages of differentiation (Figure 1b) In

other silica skeletons of basal stem the sections were

characterised by many small punctate deposits of silica,

<1μm across, while the stomata, ca 40-50 μm in

dia-meter, were more numerous, only lightly silicified and

many appeared to be linked in pairs Adjacent

epider-mal-like cells were ca 100-200μm in length and 40-50

μm wide and included highly fluorescent silica deposits

which, concomitant with their parent silica cells

appeared to be in the process of division (Figure 1c)

Some sections of silicified stem showed silica cells

which were 100-400 μm in length but without the

intracellular, nucleus/vesicle-like deposits seen in other stem sections The silicified cell walls were heavily inva-ginated and, again, included punctate and equidistant deposits of silica which as suggested previously may be indicative of the positions of plasmodesmata (Figure 1d) Silica skeletons of distal stem sections were quite differ-ent from basal sections in that they were characterised

by rosette-like accumulations of silica deposits approxi-mately 20-30 μm in diameter as well as guard cells of stomata studded with silica deposits of ca 1-2μm across and resembling ‘teeth’ where they extended into the sto-matal pore (Figure 1e) These silica rosettes appeared to

be further elaborated in nodal regions where they formed doughnut-like structures, up to ca 40μm in dia-meter, which gave the distinct impression of being silici-fied pores (Figure 1f) Other nodal regions showed long,

ca 200-500 μm, epidermal-like cells in which their jagged-in-appearance cell walls were heavily silicified There were neither punctate silica deposits nor intracel-lular silica inclusions evident in these structures (Figure 1g) The leaves showed silica skeletons which were very similar to those of the nodal regions though perhaps showing higher densities of the rosette-like silica struc-tures (Figure 1h) Stomata were heavily silicified in some sections of leaf and showed clear anatomical details including an anular ring between the pore-forming guard cells Again stomata often appeared as pairs con-nected by silicified threads of varying diameters (Figure 1i) Spores were found to be heavily silicified, being associated with spore walls and present as sub-micron punctate deposits of silica upon individual silicified spores which were between 20 and 40μm in diameter (Figure 1j) Horsetail grown from rhizomes collected from the wild under hydroponic conditions in the absence of silicic acid grew normally without any obvious requirement for silicon Acid digestion of tissues from these plants revealed no silica deposits or skeletons

PDMPO as a fluorescent indicator of silica formation in vitro

Buffer solutions at pH 7 and including 0.125 μM PDMPO showed no green fluorescence indicative of silica and only occasional particles of blue fluorescence probably due to dust or insoluble contaminants in the buffer (Figure 2a) Buffer solutions at pH 7 and includ-ing 5% w/v callose and PDMPO, but not Si(OH)4, showed no green fluorescence while callose was indi-cated as amorphous blue fluorescence (Figure 2b) Buf-fer solutions at pH 7 and including 1 mM Si(OH)4

(undersaturated) and 5% w/v callose showed significant green fluorescence in the presence of PDMPO (Figure 2c) The fluorescent material was primarily made up of aggregates of sub micron-sized particles (Figure 2c

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200 —m 200 —m

100 —m

100 —m

100 —m

100 —m

100 —m

200 —m

Figure 1 PDMPO-labelled silica deposition in horsetail a Rhizome; b Basal stem, arrows (main and insert) indicate punctate deposits of silica associated with cell walls; c Basal stem, arrow (insert) indicates silica deposition at cell plate between dividing cells; d Basal stem, arrow (insert) indicates punctate deposits of silica associated with highly invaginated cell walls; e Distal stem, showing (main and insert) rosette-like silica structures and heavily silicified stomata; f Node, showing high density of silicified structures including doughnut-like pore (insert); g Node, showing jagged appearance of silica-rich cell walls; h Leaf, showing high densities of rosette-like silica structures; i Leaf, demonstrating the intimate association of silica with stomata (insert); j Spores, showing heavily silicified spores including (insert) punctate deposits of silica on the spore surfaces Scale bars; 100 μm - d,e,f,g,h,i; 200 μm - a,b,c,j.

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insert and arrow) and these appeared to be associated

with or occluded within the blue fluorescent callose

Identical solutions in the absence of callose showed no

green fluorescence and were similar to image Figure 2a

In buffer solutions at pH 7 which included 2 mM Si

(OH)4 and 5% w/v callose the PDMPO-positive green

fluorescence was more extensive than at 1 mM Si(OH)4

and included diffuse and particulate materials, the latter

again being composed primarily of sub micron-sized

particles (Figure 2d) Identical solutions in the absence

of callose showed a significantly lesser amount of

PDMPO-positive green fluorescence and the fluorescent

material was similar in appearance and size to that

observed in the presence of callose (Figure 2e) In buffer

solutions in which the concentration of Si(OH)4 was 4

mM (saturated) there were significant flocs of

PDMPO-positive materials and particularly so in those

prepara-tions which included 5% w/v callose (Figure 2f)

The presence of silica in an undersaturated (2 mM)

solution of Si(OH)4 at pH 7 and including 5% w/v

cal-lose was further supported by fluorescence spectrometry

which demonstrated a callose-dependent shift in

emis-sion maximum from 450 to 510 nm (Figure 3a,b) That

this shift was due to the formation of silica was

con-firmed in a saturated (7 mM) solution of Si(OH)4 under

the identical solution conditions (Figure 3c) The

silica-dependent shift was significantly more pronounced in

the presence than absence of callose

Discussion When fresh or dried samples of horsetail were digested

in concentrated acid using a microwave oven all the organic materials associated with the plants were com-pletely dissolved leaving behind elaborate and detailed silica‘skeletons’ of the different plant regions The sus-pension of these silica remains in buffered solutions at

pH 7 which contained the fluorescent probe, PDMPO, enabled their detailed structures to be viewed by fluores-cence microscopy (Figure 1) It was of note that horse-tail grown hydroponically in the complete absence of added silicic acid grew normally for 10 weeks though without leaving any trace of silica following tissue diges-tion While there was no immediate evidence that horsetail required silicon for normal growth it was observed that after 10 weeks of hydroponic culture in the absence of added silicic acid some plants showed wilting and blackening of distal branch tips similar to symptoms of‘silicon-deficiency’ observed by Chen and Lewin [17] However, herein these symptoms appeared simultaneously in parts of the plants where there was evidence of infection by powdery mildew fungus and so

it was not clear as to whether they were the result of silicon deficiency or fungal infection [18] There was no evidence of fungal infection in plants grown in the pre-sence of added silicic acid While it was clear in horse-tail collected locally or grown in silicon-replete hydroponic media that silica was deposited extensively

Figure 2 PDMPO-labelled silica in vitro All [PDMPO] are 0.125 μM; All solutions are 20 mM PIPES at pH 7 All [callose] are 5% w/v a PDMPO only; b PDMPO + callose; c PDMPO + callose + 1 mM Si(OH) 4 ; the insert shows a close-up of one of the silica clusters; d PDMPO + callose + 2

mM Si(OH) 4 ; the insert shows a close-up of the precipitated silica; e PDMPO + 2 mM Si(OH) 4 ; the insert shows a close-up of silica; f PDMPO + callose + 4 mM Si(OH) 4 ; the insert shows an example of silica formed in this treatment Scale bars; 100 μm - b-f; 200 μm - a.

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throughout the stem and leaf certain structures showed

intense fluorescence which suggested significant silica

deposits in these regions Stomata were often intensley

fluorescent (Figure 1) and it was noted that silicification

of stomata in horsetail appeared to mirror the known

deposition of the hemicellulose, callose, in guard cell

differentiation and stomatal pore formation in the

related fern, Asplenium nidus [19-21] The observed

similarities between the deposition in stomatal

struc-tures of callose in A nidus and silica in E arvense were

remarkable For example, in early post cytokinetic guard

cells the nascent ventral wall was silicified (Figure 4a)

In later examples, the ventral, dorsal and periclinal walls

as well as the wall thickenings were are all silicified (Fig-ure 4b) In some stomata silicification was reduced at the centre of the ventral wall as stomatal pore formation was iniated (Figure 4c) Thereafter in further differen-tiated examples of stomata radial fibrillar arrays of silica were observed on the periclinal wall where stomatal pore formation takes place (Figure 4d) Finally in more mature stomata the wall thickenings were silicified and punctate deposits of silica were observed associated with cell walls (Figure 4e) Annular rings of silica were also observed lining the stomatal pore in more mature sto-mata (Figure 1i) All of these observations of silica deposition in E arvense have been identified as sites of callose deposition in A nidus (Figure 4) in the recent seminal and detailed studies of Apostolakos and collea-gues [19-21] These very close associations between the known deposition of callose in differentiating stomata and the presence of silica now strongly implicate callose,

or possibly, callose in conjunction with an underlying microtubule array, in directing the silicification of sto-mata in horsetail Further strong evidence that callose was involved in templating the deposition of silica else-where in horsetail was observed in silica skeletons of cells undergoing cytokinesis (Figure 5) Again silica deposition at phragmoplasts and eventually at cell plates and young cell walls dividing daughter cells mirrored the known deposition of callose in cytokinesis [22-24]

In some cells which were at an early stage of division, in some cases before there was any evidence of silica deposition at the phragmoplast, the cytosolic (and per-haps nuclear) fragments of the emerging daughter cells were found to be heavily silicified (Figure 1c) The iden-tity of these silica ‘nuclei/vesicles’ is a mystery though they may provide evidence for a role for callose in the partitioning of cytosolic and nuclear materials during cell division? The significant deposits of silica within cell walls is supported by the known presence of callose in cell walls of horsetails [12,15,25,26] In addition, equidi-stant punctate deposits of silica associated with cell walls may be indicative of, again, the known deposition

of callose in plasmodesmata (Figure 1b,d) [27,28] Finally, the heavily silicified spores (Figure 1j) may also

be evidence of the role which is known to be played by callose deposition in plant reproduction [24,29] Other silica deposits observed in horsetail may also be related

to callose deposition For example, the punctate deposits

of silica, sometimes singular and sometimes organised into rosette-like structures, which could be found throughout stem and leaf tissues were identical to those found associated with mature stomata where they are known to mimic callose deposition [30] In addition the silicified pores of internal diameter 3-5μm which were identified in leaf tissues (Figure 1f) are not dissimilar to

5% Callose + Buffer / PDMPO

Buffer/PDMPO only

510 nm

450 nm

71.2

5

-2

52.2

0.8

61.4

650

2 mM Si(OH)4+

Buffer/PDMPO

2 mM Si(OH)4+ 5% Callose + Buffer/PDMPO

7 mM Si(OH)4+ 5% Callose + Buffer/PDMPO

7 mM Si(OH)4+ Buffer/PDMPO

a)

b)

c)

Figure 3 Emission spectra (Perkin-Elmer LS50B; Ex; 338 nm;

Em: 400-650 nm) of 0.125 μM PDMPO in 20 mM PIPES

solutions at pH 7 and; a with or without 5% w/v callose; b 2 mM

Si(OH) 4 with or without 5% w/v callose; c 7 mM Si(OH) 4 with or

without 5% w/v callose.

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callose lined sieve pores found, for example, in A

thali-ana [31] We have successfully applied the fluor

PDMPO to demonstrate the deposition of silica in

horsetail and in doing so we have identified several

novel aspects of biosilicification in horsetail and in

parti-cular we have highlighted a potential role for callose in

templating silica deposition Callose biochemistry is, of

course, essential in horsetail [15,25], as in many other

plants such as the ferns [19-21], and so it is not

imme-diately evident as to how to test whether callose is

ulti-mately required for silica deposition For example,

horsetail is unlikely to grow and/or prosper if the callose

synthase gene is knocked out However, we have been

able to support our microscopy evidence linking silica

and callose deposition by demonstrating that an

under-saturated solution of Si(OH)4(i.e a solution where the

[Si(OH)4]≤ 2 mM) can be induced to form silica in the

presence of callose The formation of silica was

con-firmed by both fluorescence microscopy (Figure 2) and

fluorimetry (Figure 3) and within the usual constraints

of such original results we believe that this is the first time that an undersaturated solution of Si(OH)4 at room temperature and pressure has been induced to form silica simply by the addition of a biomolecule When silica extracted from horsetail was added to a 20

mM PIPES-buffered solution at pH 7 which included 0.125 μM PDMPO the emission spectrum changed to give a single emission maximum at ca 510 nm This positive control confirmed the known silica-induced shift in the emission spectrum of the fluor PDMPO A similar shift was also seen for solutions under the same conditions but including 5% w/v callose and either 2 or

4 mM Si(OH)4 (Figure 3) The former represents an undersaturated solution of Si(OH)4and offered up the first evidence that callose could induce Si(OH)4to auto-condense and form silica However, the in vitro evidence was most compelling in preparations containing only 1

mM Si(OH)4 when viewed by fluorescence microscopy (Figure 2c) In the absence of callose no silica could be identified by fluorescence microscopy in such

Figure 4 The deposition of callose (diagrams) and silica (fluorescent images) in the differentiation of stomata in E arvense a Callose (yellow) and silica (arrow) deposition at the nascent ventral wall (VW) of post-cytokinetic guard cells; b Deposition of callose (yellow) and silica (arrows) in the periclinal wall and dorsal wall (DW) and callose/silica remaining in the ventral wall; c Callose (yellow) and silica (arrow) disappear from the centre of the ventral wall during pore initiation; d Callose (yellow) and silica (arrows) appears as a radial fibrillar array as the stomatal pore is formed; e Upon stomatal pore formation callose (yellow) and silica (arrows) remain as punctate deposits upon the guard cell walls All stomata are ca 40 μm in diameter Information on deposition of callose taken from [19-21].

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preparations while in the presence of callose there were

clear and numerous deposits of silica some of which

were spherical and approximately 0.5 - 1.0μm in

dia-meter Intriguingly the silica bodies were intimately

associated with the polymer network of the callose,

identified as blue fluorescence, which suggested that the

constrained environment generated by the gel-like

cal-lose provided the conditions under which an

undersatu-rated solution of Si(OH)4(1 mM) could be‘tricked’ into

undergoing autocondensation and subsequent growth

towards stable aggregates of silica Callose is a linear

homopolymer made up primarily ofb-1,3-linked glucose

residues which at the concentration used herein, ca 5%

w/v, will form a viscoelastic gel [32] within which the

orientation of hydroxyl groups on the glucose

mono-mers may be such that they are able to iniate the first

steps in the autocondensation of silicic acid as it slowly

diffuses within the callose matrix The hydroxyl groups

on the polymer network of callose in some way enable

the energy barrier to the autocondensation of Si(OH)4

to be overcome and once the first Si-O-Si linkages have

been made further condensation reactions can proceed

much more easily to eventually build the silica

aggre-gates observed, for example, in Figure 2c While further

experiments will be required to delineate the range of

conditions under which callose induces silica formation

in undersaturated solutions of Si(OH)4 and the exact

mechanism by which this is achieved we now have a

long sought after biomolecule which can act as a

tem-plate for silica formation and deposition in vitro If this

is also the basis for the mechanism of silica deposition

in horsetail then it may also be significant in other lose producing biosilicifiers such as diatoms [33] If cal-lose is the key then associated biochemistry including enzymes such as callose synthase (potentially catalysing Si-O-Si bond formation) and b-1,3-glucanases (poten-tially cleaving Si-O-Si bonds) [25] will play a pivotal role

in the modelling and remodelling of silica frameworks The deposition of silica in horsetail has been studied for many decades and we now have a possible mechanism

of silica deposition in this plant which could also be a general mechanism of biosilicification

Conclusion The fluor PDMPO has been used to identify silica deposition in horsetail and to provide new insight into silicification in this plant It was observed that silica deposition in horsetail exactly mirrored the known deposition of callose in the related fern and other plants Callose was shown to induce the formation and precipi-tation of silica in undersaturated solutions of silicic acid This was the first time that this had been demonstrated for any biomolecule and it suggested that callose and perhaps other similar carbohydrates might be key mole-cules in biological silicification

Methods

Hydroponic culture of horsetail

Horsetail (Equisetum arvense) rhizomes were collected locally, washed in ultrapure water (conductivity < 0.067 μS/cm) and subjected to hydroponic culture in 1/6th

MS medium in the presence (2 mM) or absence of added silicic acid The latter media included an additional 8

mM Na+ to account for Si addition as Na4SiO4 After 10-12 weeks of a 14 h light/10 h dark cycle at 25°C healthy horsetail plants had grown under both sets of conditions

Digestion of horsetail materials

Horsetail plants, either collected locally or grown hydro-ponically, were washed in ultrapure water, allowed to air-dry, cut into discrete 1 cm sections of rhizome/root, basal stem, distal stem, nodal regions and leaves and ca 0.5 g of each placed in acid-washed 20 mL PFA teflon© vessels The samples were then digested in a 1:1 mixture

of 15.8M HNO3 and 18.4M H2SO4 using a Mars Xpress microwave oven (CEM Microwave Technology Ltd.) The acid digests were clear and, upon dilution with 8

mL of ultrapure water, were filtered and the residues washed several times with further volumes of ultrapure water Filters were then placed in plastic Petri dishes in

an incubator at 37°C to achieve dryness over several days Dry residues off each filter were then collected into Bijoux tubes and stored in a dry, sealed, perspex cabinet

a)

b)

Figure 5 a,b PDMPO-labelling of silica deposition of cell plates

and young cell walls (arrows) forming in cytokinetic cells.

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PDMPO labelling of horsetail silica

Silica residues collected from filters were suspended in

20 mM PIPES at pH 7 and containing 0.125 μM

2-(4-pyridyl)-5-((4-(2-dimethylaminoethylaminocarbamoyl)

-methoxy)phenyl)oxazole (PDMPO; LysoSensor Yellow/

Blue DND-160, 1 mM in DMSO) This intracellular pH

probe [34] has been shown to be bound by silica (but

not silicic acid) and to emit ‘green’ fluorescence upon

excitation at 338 nm [35-38] Suspensions were left for

24 h to allow the reaction between silica surfaces and

PDMPO to achieve completion after which 50μL

sam-ples were transferred to a cavity slide and viewed using

an Olympus BX50 fitted with a BXFLA fluorescent

attachment using a U-MWU filter cube (Ex: 333-385

nm; Em: 400-700 nm) A ColourView III digital camera

(OSIS FireWire Camera 3.0 digitizer) was used to

cap-ture images in conjunction with CELL* Imaging

soft-ware (Olympus Cell* family, Olympus Soft Imaging

solutions GmbH 3.0)

In vitro preparations of callose and silicic acid

Callose (b-D Glucan, Barley, Sigma, UK) was dissolved

at 5% w/v in 20 mM PIPES buffer solutions at pH 7 and

containing 0, 1, 2, 4 and 7 mM Si(OH)4 by warming

each preparation in a water bath at 100°C for 60

sec-onds Upon cooling to room temperature PDMPO was

added to a concentration of 0.125 μM Equivalent

con-trol solutions to which no callose had been added were

treated in an identical manner All solutions were then

incubated at room temperature in the dark for 5 days

before being examined by fluorescence microscopy, see

above, or their emission spectra were determined by

fluorimetry (Perkin-Elmer LS50B; Ex; 338 nm; Em:

400-650 nm) as previously described [35]

Acknowledgements

CL was in receipt of a NERC studentship.

Authors ’ contributions

CE designed the study and provided training and guidance in experimental

methods CE wrote and prepared the first draft of the manuscript CL carried

out the majority of the experimental work and helped with writing the

manuscript.

Both authors have read and approved this manuscript.

Competing interests

The authors declare that they have no competing interests.

Received: 15 April 2011 Accepted: 29 July 2011 Published: 29 July 2011

References

1 Exley C: Silicon in life: a bioinorganic solution to bioorganic essentiality J

Inorg Biochem 1998, 69:139-144.

2 Exley C: Darwin, natural selection and the biological essentiality of

aluminium and silicon Trends Biochem Sci 2009, 34:589-593.

3 Epstein E: The anomaly of silicon in plant biology Proc Natl Acad Sci USA

1994, 91:11-17.

4 Currie HA, Perry CC: Silica in plants: biological, biochemical and chemical

5 Exley C: Silicon in life: whither biological silicification? In Biosilica in Evolution, Morphogenesis and Nano-biotechnology Edited by: Muller WEG, Grachev MA Springer; 2009:173-184.

6 Page CN: An assessment of inter-specific relationships in Equisetum subgenus Equisetum New Phytol 1972, 71:355-369.

7 Kaufman PB, LaCroix JD, Dayanandan P, Allard LF, Rosen JJ, Bigelow WC: Silicification of developing internodes in the perennial scouring rush (Equisetum hyemale var affine) Developmental Biol 1973, 31:124-135.

8 Laroche J, Guervin C, Le Coq C, Robert D: Activités pétrogénétiques chez Equisetum arvense L (Ptéridophytes) Bulletin de la Société Botanique de France 1992, 139:47-55.

9 Holzhüter G, Narayanan K, Gerber T: Structure of silica in Equisetum arvense Analyt Bioanalyt Chem 2003, 376:512-517.

10 Sapei L, Nöske R, Strauch P, Paris O: Isolation of mesoporous biogenic silica from the perennial plant Equisetum hyemale Chem Mat 2008, 20:2020-2025.

11 Gierlinger N, Sapei L, Paris O: Insights into the chemical composition of Equisetum hyemale by high resolution Raman imaging Planta 2008, 227:969-980.

12 Currie HA, Perry CC: Chemical evidence for intrinsic ‘Si’ within Equisetum cell walls Phytochemistry 2009, 70:2089-2095.

13 Perry CC, Fraser MA: Silica deposition and ultrastructure in the cell wall of Equisetum arvense: the importance of cell wall structures and flow control in biosilicification? Phil Trans Roy Soc London B 1991, 334:149-157.

14 Perry CC, Lu Y: Preparation of silicas from silicon complexes: Role of cellulose in polymerisation and aggregation control J Chem Soc Faraday Trans 1992, 88:2915-2921.

15 Fry SC, Nesselrode BHWA, Miller JG, Mewburn BR: Mixed linkage (1 →3,1→4)-β-glucan is a major hemicellulose of Equisetum (horsetail) cell walls New Phytol 2008, 179:104-115.

16 Perry CC: An overview of silica in biology: Its chemistry and recent technological advances In Biosilica in Evolution, Morphogenesis and Nano-biotechnology Edited by: Muller WEG, Grachev MA Springer; 2009:295-313.

17 Chen CH, Lewin J: Silicon as a nutrient element for Equisetum arvense Can J Bot 1969, 47:125-131.

18 Fauteux F, Chain F, Belzile F, Menzies JG, Belanger RR: The protective role

of silicon in the Arabidopsis-powdery mildew pathosystem Proc Natl Acad Sci USA 2006, 103:17554-17559.

19 Apostolakos P, Livanos P, Galatis B: Microtubule involvement in the deposition of radial fibrillar callose arrays in stomata of the fern Asplenium nidus L Cell Motility Cytoskeleton 2009, 66:342-349.

20 Apostolakos P, Livanos P, Nikolakopoulou TL, Galatis B: The role of callose

in guard-cell wall differentiation and stomatal pore formation in the fern Asplenium nidus Ann Bot 2009, 104:1373-1387.

21 Apostolakos P, Livanos P, Nikolakopoulou TL, Galatis B: Callose implication

in stomatal opening and closure in the fern Asplenium nidus New Phytol

2010, 186:623-635.

22 Scherp P, Grotha R, Kutschera U: Occurrence and phylogenetic significance of cytokinesis-related callose in green alga, bryophytes, ferns and seed plants Plant Cell Rep 2001, 20:143-149.

23 Thiele K, Wanner G, Kindzierski V, Jürgens G, Mayer U, Pachl F, Assaad FF: The timely deposition of callose is essential for cytokinesis in Aribodopsis The Plant J 2009, 58:13-26.

24 Chen XY, Kim JY: Callose synthesis in higher plants Plant Signall Behav

2009, 4:489-492.

25 Fry SC, Mohler KE, Nesselrode BHWA, Franková L: Mixed linkage β-glucan: xyloglucan endotransgluocosylase, a novel wall remodelling enzyme from Equisetum (horsetail) and charophytic algae The Plant J 2008, 55:240-252.

26 Sørensen I, Pettolino FA, Wilson SM, Doblin MS, Johansen B, Bacic A, Willats WGT: Mixed linkage (1 →3, 1→4)- β-D-glucan is not unique to the Poalesm and is an abundant component of Equisetum arvense cell walls The Plant J 2008, 54:510-521.

27 Xu XM, Jackson D: Lights at the end of the tunnel: new views of plasmodesmal structure and function Current Opin Plant Biol 2010, 13:684-692.

28 Zavaliev R, Ueki S, Epel BL, Citovsky V: Biology of callose ( β-1,3-glucan) turnover at plasmodesmata Protoplasma 2011, 248:117-130.

29 Nishikawa SI, Zinkl GM, Swanson RJ, Maruyama D, Preuss D: Callose (beta-1,3-glucan) is essential for Aribodopsis pollen wall patterning, but not tube growth BMC Plant Biol 2005, 5:art 22.

Trang 9

30 Majewska-Sawka A, Münster A, Rodríguez-García MI: Guard cell wall:

immunocytochemical detection of polysaccahride components Journal

of Experimental Botany 2002, 53:1067-1079.

31 Thompson MV, Wolniak SM: A plasma membrane-anchored fluorescent

protein fusion illuminates sieve element plasma membranes in

Aribodopsis and tobacco Plant Physiol 2008, 146:1599-1610.

32 Cui SW, Wang Q: Cell wall polysaccharides in cereals: chemical structures

and functional properties Struct Chem 2009, 20:291-297.

33 Størseth TR, Kirkvold S, Skjermo J, Reitan KI: A branched β-D-(1→3,

1→6)-glucan from the marine diatom Chaetoceros dibilis (Bacillariophyceae)

charcaterised by NMR Carbohydrate Res 2006, 341:2108-2114.

34 Diwu Z, Chen CS, Zhang C, Klaubert DH, Haugland RP: A novel acidotropic

pH indicator and its potential application in labeling acidic organelles of

live cells Chem and Biol 1999, 6:411-418.

35 Shimizu K, Del Amo Y, Brzezinski MA, Stucky GD, Morse DE: A novel

fluorescent silica tracer for biological silicification studies Chem Biol

2001, 8:1051-1060.

36 Leblanc K, Hutchins DA: New applications of a biogenic silica deposition

fluorophore in the study of eceanic diatoms Limnol Oceanography::

Methods 2005, 3:462-476.

37 Hazelaar S, van der Strate HJ, Gieskes WWC, Vrieling EG: Monitoring rapid

valve formation in the pennate diatom Navicula salinarum

(Bacillariophyceae) J Phycol 2005, 41:354-358.

38 Ogane K, Tuji A, Suzuki N, Kurihara T, Matsuoka A: First application of

PDMPO to examine silicification in polycystine Radiolaria Plankton

Benthos Res 2009, 4:89-94.

doi:10.1186/1471-2229-11-112

Cite this article as: Law and Exley: New insight into silica deposition in

horsetail (Equisetum arvense) BMC Plant Biology 2011 11:112.

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